Rahaman-Noronha E, Donnell M, Pilley C, Wright P
J Exp Biol. 1996;199(Pt 12):2713-23. doi: 10.1242/jeb.199.12.2713.
This study examined ammonia excretion by embryos of the rainbow trout (Oncorhynchus mykiss). The distribution of ammonia in relation to the H+ distribution and electrical potential was determined. The influence of the pH of the unstirred layer (USL) of water next to the external surface of the embryo was also assessed. Eyed-up embryos (3540 days post-fertilization) were exposed to various external water conditions [pH 6.0, pH 10.0, 1.6 mmol l-1 NaCl, 0.0 mmol l-1 NaCl, 0.2 mmol l-1 NH4Cl, 2.5 mmol l-1 borax buffer (Na2B4O7.10H2O), 2.5 mmol l-1 Hepes, 0.1 mmol l-1 amiloride] for 30 min and ammonia excretion rates, ammonia concentration in the perivitelline fluid (PVF) and yolk, and the pH of the PVF, yolk and USL were measured. The rate of ammonia excretion was dependent, in part, on the partial pressure gradient of NH3 ( PNH3) from the PVF to the USL. Exposure to water of pH 6 increased, whereas NH4Cl or pH 10 exposure decreased, ammonia excretion rates. Elevated external Na+ levels also influenced the rate of ammonia excretion, but neither Na+-free water nor amiloride had any effect. The distribution of ammonia between the PVF and USL was dependent on the H+ distribution, but ammonia was distributed according to the electrical potential between the PVF and yolk. The USL was 0.32 pH units more acidic than the bulk water. Addition of buffer to the external water eliminated the acid USL and decreased ammonia excretion rates. We conclude that rainbow trout embryos excrete ammonia primarily as NH3, but when external Na+ levels are elevated, ammonia excretion may be independent of the PNH3 gradient. The acidic USL next to the chorion probably facilitates NH3 diffusion by maintaining the PNH3 through the conversion of NH3 to NH4+ upon entry into the USL.
本研究检测了虹鳟(Oncorhynchus mykiss)胚胎的氨排泄情况。确定了氨与H⁺分布及电势相关的分布情况。还评估了胚胎外表面附近水的未搅动层(USL)的pH值的影响。将眼点形成的胚胎(受精后35 - 40天)暴露于各种外部水条件下[pH 6.0、pH 10.0、1.6 mmol l⁻¹ NaCl、0.0 mmol l⁻¹ NaCl、0.2 mmol l⁻¹ NH₄Cl、2.5 mmol l⁻¹硼砂缓冲液(Na₂B₄O₇·10H₂O)、2.5 mmol l⁻¹ Hepes、0.1 mmol l⁻¹氨氯吡咪] 30分钟,并测量氨排泄率、卵周液(PVF)和卵黄中的氨浓度以及PVF、卵黄和USL的pH值。氨排泄率部分取决于从PVF到USL的NH₃分压梯度(PNH₃)。暴露于pH 6的水中会增加氨排泄率,而暴露于NH₄Cl或pH 10的水中会降低氨排泄率。外部Na⁺水平升高也会影响氨排泄率,但无Na⁺水和氨氯吡咪均无任何影响。PVF和USL之间氨的分布取决于H⁺分布,但氨是根据PVF和卵黄之间的电势分布的。USL比主体水的酸性高0.32个pH单位。向外部水中添加缓冲液可消除酸性USL并降低氨排泄率。我们得出结论,虹鳟胚胎主要以NH₃形式排泄氨,但当外部Na⁺水平升高时,氨排泄可能与PNH₃梯度无关。绒毛膜附近的酸性USL可能通过在NH₃进入USL时将其转化为NH₄⁺来维持PNH₃,从而促进NH₃扩散。